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  • Articles  (199)
  • Cell & Developmental Biology  (199)
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  • 101
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 81 (1973), S. 133-138 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Balb/3T3 cells show density-dependent regulation of multiplication with the final cell density depending on serum concentration in the media. Chemically transformed Balb/3T3 cells (Balb/3T3-D) pile up on each other, multiply to a high cell density, but have decreased DNA synthesis at very high cell densities. Balb/3T3-D cells require less serum for multiplication compared with original Balb/3T3 cells. A rat serum fraction and a bovine β-globulin fraction stimulate the multiplication of Balb/3T3 cells but only slightly stimulate Balb/3T3-D cells indicating different serum factors stimulate growth of these two cell types. The multiplication properties of Balb/3T3-D cells are very similar to those of SV-40 transformed 3T3 cells, however, these properties were brought about by a single treatment by a chemical carcinogen, without an exogenous virus. The transformation altered the contact of cells to one another, indicating a permanent chemical change in the membrane structure.
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  • 102
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 81 (1973), S. 149-151 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Warming of exponentially growing T. pyriformis to 34°C results in severe inhibition of nucleotide pool formation. The utilization of the pool for stable RNA synthesis is poorly affected at the high temperature. It thus appears that the synthesis and processing of ribosomal RNA precursors are not primarily impaired at 34°C.
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  • 103
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 81 (1973), S. 171-180 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Rates of incorporation of labelled thymidine (RIT), radioautographic labelling index for DNA synthesis (LI) and mitotic incidence following colcemid metaphase arrest (MI) were measured in organ cultures of newborn and adult rat lung. In adult cultures these three parameters correlated well, being low after explantation and reaching a maximum after two to three days. In newborn cultures RIT fell several fold over the first 24 hours after explantation and, in this respect, did not correlate with LI and MI. The changes in RIT over the first 24 hours appear to be due to changes in the degree of competition between endogenous TdR and exogenous labelled TdR, probably caused by leakage of the intracellular thymidine pool following explantation. The report emphasizes the need to check RIT data against radioautographic evidence before accepting it as an index of DNA synthesis.
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  • 104
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The Rapidly Migrating Proteins (RMP) which shuttle nonrandomly between nucleus and cytoplasm and equilibrate in approximately equal amounts in each compartment, were isolated from Amoeba proteus by implanting 3H-protein containing nuclei into unlabeled cells and some time later extracting the labeled material from the cytoplasms of such cells. The labeled material was subsequently fractionated by gel filtration in Sephadex G-100 columns. The RMP are soluble in dilute salt solutions and appear as a heterogenous group of molecules, one component of which seems to be a single species of protein accounting for ca. one-third of the RMP fraction. Because of its distinctness this component, called the LR fraction, received the major attention in this study. LR was found to comprise ca. 17% of the aqueous-soluble proteins of the nucleus and ca. 3-4% of the total cell protein.LR has a very low molecular weight as determined, e.g., by its elution from a Sephadex G-100 column. Because of its low molecular weight, LR could be purified by taking advantage of the fact that LR is (1) soluble in a saturated Solution of ammonium sulfate and (2) insoluble in butanol, diethyl ether, and 10% trichloroacetic acid.LR migrates toward the anode as a single band when subjected to electrophoresis on “standard disc” and SDS polyacrylamide gels. It does not enter a gel designed to separate basic proteins (at pH 4.0). When subjected to Sephadex G-25 gel filtration LR migrates through the gel as a single band and elutes from the gel at a position in the middle of the linear separation range that indicates its molecular weight is ca. 2300. The only N-terminal amino acid found in the LR fraction is proline.Evidence is presented to show that LR is not the product of a non-specific breakdown of protein produced during its isolation, but the possibility that it results from the cleavage of a single chemical bond of a larger polypeptide, has not been eliminated.When injected into non-labeled amebae, purified radioactive LR concentrates in the nucleus  -  just as radioactive RMP concentrates in a recipient cell nucleus when an amino acid-labeled nucleus is implanted into an unlabeled cell.
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  • 105
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 82 (1973), S. 511-512 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 106
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 81 (1973), S. 113-123 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Fetal rat hepatocytes in primary monolayer cultures multiply in arginine-deficient medium. Both the “recovery efficiency” and the final cell density of the cultured cells are proportional to the concentration (0-15%, v/v) of dialyzed fetal bovine serum in the medium. Stationary-phase cells divide again following addition of fresh serum to the culture. After two to three generations of growth, the chromosome number of these cells remains diploid [2N = 42].Cross-feeding (of a subpopulation of arginine-requiring liver-derived cells by parenchymal arginine-synthesizing cells) and cellular degradation of various serum proteins do not account for sources of arginine required for cell multiplication in this culture system. Because these cultured hepatocytes utilize ornithine for arginine biosynthesis, and because ornithine enhances the rate and the amount of cell multiplication, it is more likely that the multiplying cells are parenchymal arginine-synthesizing hepatocytes.At least two classes of serum factors are required for the growth of cultured fetal rat hepatocytes: one stimulates cell multiplication; the other is required for cellular survival and/or attachment to the culture dish. These factors have been partially separated by fractionation with ammonium sulfate; they are non-dialyzable, heat labile, and sensitive to changes in pH.
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  • 107
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 81 (1973), S. 161-170 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A partially purified multiplication-stimulating activity for chicken embryo fibroblasts in cell culture was isolated from rat liver cell conditioned medium (see preceding paper, Dulak and Temin, 1973). It has been analyzed by isoelectric focusing and by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Multiplication-stimulating activity resided in a family of at least four polypeptides which were similar in apparent molecular size, but different in electrical charge. These polypeptides have a specific activity of about 50,000 with respect to serum. One of them has been purified on a small scale to apparent homogeneity in a sodium dodecyl sulfate polyacrylamide gel.
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  • 108
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    Journal of Cellular Physiology 81 (1973), S. 217-224 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Transplantable SV40-transformed hamster cells cultivated in the presence of low concentrations of BrdU for prolonged periods of time and cells made deficient in the enzyme thymidine kinase (dTK) by continued exposure to BrdU became less tumorigenic. In both instances, when grown in BrdU the cells contained analog substituted DNA. The tumorigenicity of dTK+ cells exposed to low concentrations of BrdU, but not the dTK- cells, returned to control values when the cells were grown in medium devoid of BrdU. A tumorigenic mouse cell line made dTK deficient also had diminished oncogenicity. However, transformed hamster cells made deficient in another salvage pathway enzyme, hypoxanthineguanine phosphoribosyl-transferase by growth in eight azaguanine, retained their tumorigenicity. Two of five revertant cell lines, in which thymidine kinase activity was restored, transplanted more readily to hamsters than the dTK- cells from which they were derived. It is concluded that there is a relative loss of tumorigenicity when BrdU is incorporated into the DNA of tumorigenic cell lines, or when there is a genetic modification of thymidine kinase activity.
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  • 109
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Quite a number of phenomena having to do with cells' influences upon one another's movements have come to be regarded as expressions of “contact inhibition.” However, no single, central mechanism has been shown to underlie them all. Consequently, the term “contact inhibition” should not be used without operational modifiers. Inhibitions of individual cell movements imputed to be mediated by cell-cell contacts include inhibition of overlapping (which results in monolayering), of colony expansion, of cell speed (nuclear translocation), of ruffling, of orthogonal movement (proposed to explain spontaneous parallel alignment of cells), and of neighbor exchanges.The six inhibitions listed above are operationally distinct, and only two (overlapping and colony expansion) are known to result from a common mechanism. A seventh phenomenon, so-called “contact inhibition of cell division” (more operationally termed postconfluence inhibition of cell division) is in a separate category and is not considered here.Evidence eliminating action-at-a-distance is available only for the first three, and hence only these should at present be termed contact inhibitions. Inhibition of neighbor exchanges is yet hypothetical; at its extreme, it would immobilize cells in a confluent monolayer, but such immobilization has been found not to occur.Contact inhibition of overlapping, the most studied of the six, is not displayed by invasive cells with respect to normal cells; invasive tumor cells overlap freely upon normal cells, although not necessarily upon one another. Contact inhibition of overlapping, and its loss by invasive cells, can readily be interpreted, by means of the differential adhesion hypothesis, as consequences of cell-type-specific differences in cell-cell and cell-substratum “strengths of adhesion.” These strengths of adhesion are formulated as specific interfacial free energies, which are the only parameters of cellular adhesiveness that have been rigorously shown to determine equilibrium configurations of cell populations.
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  • 110
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    Journal of Cellular Physiology 81 (1973), S. 85-89 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The embryonic chick skeletal muscle cells differentiated in cell culture from trypsin-dissociated myoblasts produce a spike response which is tetrodotoxin-sensitive. It has been found that many cells also produce a plateau response which is resistant to tetrodotoxin. The plateau response frequently occurs even in the muscle cells which do not normally exhibit the spike response. During the plateau response membrane resistance is greatly reduced below its resting value. The current-voltage relation in muscle cells with the plateau response is always S-shaped. It is suggested that the plateau arises from a voltage-dependent increase in permeability to external cations whose influx produce the maintained depolarization, and from low level of repolarizing potassium outflux. The plateau response is sensitive to manganese ions. This finding, together with resistibility to tetrodotoxin, suggests that calcium ions are the dominant carriers for the depolarizing current.
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  • 111
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Aminonucleoside (AMS) inhibited the cell cycle of human lung fibroblasts at a point in G1 phase and at another point in G2 phase. Even when this inhibition was fully established, DNA synthesis and mitosis which were in progress proceeded normally. Inhibition of RNA synthesis in the cultures preceded the effects on DNA synthesis and mitosis, but inhibition of protein synthesis could not be detected.These points of potential inhibition do not exist in the cell cycle of HeLa cell, or are not affected by aminonucleoside. Here inhibition of cell proliferation by AMS was less marked, and when inhibition eventually occurred it was not specific for any point of the cell cycle. The rate of entry of the inhibitor was similar in both types of cell.
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  • 112
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 81 (1973), S. 125-131 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Adenosine (10 μM) stimulates the initial growth rate of BHK/21 cells seeded at low but not high density in monolayer culture; it does not affect final cell density or permit growth in agar. In labelling experiments With tritiated thymidine, adenosine also increases the response of quiescent cells to low concentrations of serum. Dialysis of serum to remove oxypurines only marginally reduces its effect on quiescent cell labelling or growth, indicating that BHK/21 cells are able to synthesise purines. The response of quiescent cells to 5% serum is inhibited by high MW (2 × 106) dextran sulphate at 2 μg per milliliter. Low MW dextran sulphate (30,000) and heparin at 20 μg per milliliter produce the same effect. Exogenous adenosine (10 μM) prevents this inhibition. Many other purine derivatives replace adenosine for all the above activities but xanthine is completely inactive in all. It, therefore, appears that nucleotide synthesis is a necessary function of these compounds.The growth of cells of a polyoma-virus-transformed BHK/21 line in monolayer is not stimulated by exogenous purine, though their colony-forming ability in agar is increased five-fold. The stimulating effects of exogenous purines on normal BHK/21 cells and the absolute requirement for them in the presence of polyanions is discussed in relation to possible mechanisms of growth control.
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  • 113
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 81 (1973), S. 153-160 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A polypeptide fraction with multiplication-stimulating activity for chicken and rat embryo fibroblasts was partially purified from serum-free medium conditioned by the growth of a line of rat liver cells. The specific multiplication-stimulating activity of this fraction was 27,000 times that of serum. The rat liver cell multiplication-stimulating activity had a molecular weight of approximately 10,000 daltons and was inactivated by mercaptoethanol and dithiothreitol. It had sulfation factor and non-suppressible insulin-like activities, but did not have anti-trypsin activity. The rat liver cell multiplication-stimulating activity resembled both multiplication-stimulating activity from calf serum and somatomedin.
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  • 114
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    Journal of Cellular Physiology 81 (1973), S. 199-216 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The isolation of a temperature sensitive cell line from the Chinese hamster line CCL39 of the American Type Culture Collection is described. At the nonpermissive temperature (39°C) the cells become attached to the surface of tissue culture dishes, but no microscopically observable colonies are formed upon prolonged incubation. Exposure to the high temperature for more than 24 hours leads to an almost complete loss in viability. A karyotypic analysis showed that this new line has lost one of the medium-sized metacentric chromosomes, although no proof is available so far to show that this loss is not simply coincidental.In nonsynchronized cultures transferred to 39°C DNA synthesis stops first, RNA synthesis shortly thereafter, while protein synthesis (turnover) continues for a longer time. After such a shift the cell number increases by less than 15% as measured with the Coulter counter.Studies with synchronized cultures give the following results: (1) one round of DNA synthesis can occur at 39°C when the cells are released from serum starvation or a hydroxyurea block, or when mitotic cells are placed at 39°C; (2) the entry of cells into metaphase of mitosis at 39°C is almost normal when the preceding time interval at 39°C is only eight hours (release of cells from G1/S boundary), but considerably reduced when the cells spend an additional 12 to 15 hours at 39°C in G1 (release from serum starvation).Infection by SV40 virus temporarily induces DNA synthesis after it has come to a stop at the nonpermissive temperature, but cells permanently transformed by SV40 still exhibit the temperature-sensitive phenotype.
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  • 115
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    Journal of Cellular Physiology 81 (1973), S. 233-239 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Calf thymus histones and histone subfractions were added to media overlying subconfluent mouse fibroblast cells in culture. The histones caused significantly higher cell densities at confluence than control cultures and disruption of the normal ordered arrangement of cells. These changes were seen on application of histones to growing cells but not confluent cells and were reversed when the histones were removed and the cells replated with more growth area. The slightly lysine rich histone fraction had the greatest effect and the lysine rich fraction had the least effect on cell morphology and cell number at confluence. These effects could not be duplicated with other highly charged basic proteins.
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  • 116
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    Journal of Cellular Physiology 81 (1973), S. 271-279 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Trypsinized cells of newborn mouse cerebellum have been separated by velocity sedimentation at unit gravity in shallow gradients of Ficoll. The two main technical difficulties were formation of gels around the dissociated cells and clumping of cells before and during the sedimentation procedure. These were solved by adding DNase to the dissociation medium and with holding serum, respectively. Proliferating cells of the external granular layer separated according to size differences in the cell generation cycle. Identification of Purkinje or other early-forming neurons was made by labeling them with 3H-thymidine on their birthdays. Many of the fractions contain viable cells capable of aggregating in culture.
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  • 117
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    Journal of Cellular Physiology 82 (1973), S. 285-297 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Intracellular measurements were made on immature cultured rat neurons using single or double barrelled microelectrodes; 361 dorsal root ganglion, DRG, cells, 105 spinal cord cells and 12 cerebellar cells were studied. Membrane potentials recorded were in the range -25 mV to -50 mV. A statistically significant increase in membrane potentials with time in culture was found for DRG cells during 32 days in culture. This was not found for spinal cord cells.Voltage-current curves showed a non-linearity in about 50% of DRG and spinal cord cells similar to anomalous rectification. Measurements made in DRG and spinal cord cells at ± 10 nA showed significantly greater average cell input resistance during hyperpolarising pulses than during depolarising pulses.The calculated values of Rm and Cm for DRG and spinal cord cells were similar in magnitude to values given for other cells by other workers. The value for L, the dimensionless electrotonic length, was slightly higher than that calculated for motoneurones by other workers. Differences between these results and those of others working on neurons in vitro are ascribed to the immature nature of the cells studied here.
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  • 118
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    Journal of Cellular Physiology 82 (1973) 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 119
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    Journal of Cellular Physiology 82 (1973), S. 333-338 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Viability, DNA synthesis and mitosis have been followed in the temperature sensitive Chinese hamster cell mutant K12 under permissive and non-permissive conditions. On incubation at 40°C cells retained their ability to form colonies at 33°C for 15 to 20 hours, but viability was lost gradually during the following 20 hours. When random cultures of K12 were shifted to 40°C the rate of DNA synthesis was normal for three to four hours but then decreased markedly, reaching 95% inhibition after 24 hours. Under the same conditions mitosis was inhibited after 15 hours. If cultures which had been incubated at 40°C for 16 hours were placed at 33°C the rate of DNA synthesis increased five hours after the shift down and mitosis 18 hours after. These results can be interpreted on the assumption that K12 at 40°C is unable to complete a step in the cell cycle which is essential for DNA synthesis and which occurs three to four hours before the start of S at 33°C.
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  • 120
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The endogenous respiration of isolated rat liver parenchymal cells prepared by two enzymatic techniques has been measured. Despite the production of a high percentage of viable cells by both methods, as measured by the exclusion of trypan blue, the cells obtained by one of the techniques consume oxygen at only one-half the rate of those obtained by the other technique. It is suggested that the lower rate is the result of an insufficient oxygen supply and also a lack of control of the pH of the incubation medium during the preparation of the cells.
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  • 121
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    Journal of Cellular Physiology 82 (1973), S. 421-433 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The initial rates of deoxy-D-glucose transport by cultures of growing and density-inhibited mouse embryo cells and lines of mouse cells transformed spontaneously or after infection by murine leukemia virus or murine sarcoma virus were investigated as a function of the deoxyglucose concentration. The apparent Km for deoxyglucose transport was about the same for all types of cells (1-2 mM). The Vmax of secondary cultures of mouse embryo cells decreased from 6 nmoles/106 cells/minute for sparse cultures to less than 1 nmole/106 cells/minute for density-inhibited cultures. The Vmax was about the same whether estimated in monolayer culture or in suspensions of cells dispersed by treatment with trypsin. The Vmax for deoxyglucose transport by the established cells, whether transformed spontaneously or by virus infection, was 4 to 25 times higher than that for density-inhibited mouse embryo cells and was independent of the cell density of the cultures. Deoxyglucose transport was competitively inhibited by Cytochalasin B, Persantin, glucose and 3-O-methyl-D-glucose and the apparent Ki values of inhibition were similar for the mouse embryo cells and the various cell lines. Similarly, the sensitivity of the glucose transport systems to inactivation by p-chloromercuribenzoate was about the same for all types of cells. The results suggest that the glucose transport system of the normal mouse embryo cells and the cells of the various established lines is qualitatively the same, but that the number of functional transport sites differs for the various cell lines and decreases markedly in mouse embryo cells with an increase in cell density of the cultures.
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  • 122
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cultures of human diploid fibroblasts (HDFs) exhibiting density dependent inhibition of replication (DDIR) resumed their progression through the cell cycle following medium replacement and, after a lag period of two hours, showed a dramatic increase in the incidence of isonucleolinar 4 cells and in the levels of uptake of 3H-uridine into the nucleoli. Between five and ten hours after refeeding these nucleolar changes were maximal, leveling off at the highest values, in periods corresponding to late G1 and early S. Concomitantly, a parallel increase in the number of nucleolini per cell occurred. As cells progressed through S and G2 phases the nucleolini decreased in number and reverted to the aniso-nucleolinar type. The intensity of nucleolar labeling by 3H-uridine and its correlate, the frequency of cells with labeled nucleoli, also decreased during these cell cycle stages. Both pre- and postreplicative periods of mitotic quiescence were characterized by high levels of anisonucleolinosis (60-80% of the cells) and by very low levels of nucleolar 3H-uridine incorporation.The magnitude of these nucleolar changes occurring during G1 stage was found to be strongly dependent on: (1) the length of time of contact between the cells and the fresh medium, at least eight hours of contact being necessary for a maximal response; (2) the amount of serum in the medium, the optimal serum concentration being between 10 and 50%, and (3) the pH of the medium. The nucleolar response was completely abolished at pH values below 7.0. These nucleolar changes were very sensitive to the presence of cycloheximide (10 μg/ml) and actinomycin D (0.003 μg/ml). The behavior of the nucleoli in response to these parameters was similar to the activation response of the cells to initiate DNA synthesis.During the time period of maximal nucleolar (activation) the onset of DNA synthesis as well as the morphological and autoradiographic manifestations of the nucleolar activation were completely inhibited by very low levels of actinomycin D (Ellem and Mironescu, '72), a selective inhibitor of nucleolar RNA synthesis (Perry, '65). This suggested a possible role of nucleolar metabolism, in normal diploid cells, in the initiation of DNA synthesis. Our results, however, seem to indicate that the nucleolar changes are necessary but not sufficient for the subsequent initiation of DNA synthesis, since with graded serum concentrations or medium volumes, smaller levels of a stimulus were needed to produce maximal isonucleolinosis than to effect a maximum replicative response in the cells.
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  • 123
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    Journal of Cellular Physiology 82 (1973), S. 489-495 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Visible light of moderate intensity (1200 ft-cd) can severely inhibit cell division of a non-photosynthetic mutant of Euglena gracilis when growth is supported by butanol, ethanol, or fumarate as sole carbon source. The degree of inhibition is pH dependent, being greatest at pH 4 to 5. A wide variety of other carbon sources permitted growth in the light without inhibition.
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  • 124
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    Journal of Cellular Physiology 82 (1973), S. 497-510 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The study of long-term cultures of myogenic cells has proven that electrical excitability develops only after the development of electrical coupling between the cells. That is, neither surface contact in itself nor coupling in itself is sufficient to cause excitability to develop in these cells. Following the formation of multinucleated myotubes, several different types of electrical responses develop. Some of the action potentials are sodium-dependent and are blocked by tetrodotoxin (TTX). Others are dependent upon sodium and possibly calcium and they are not blocked by TTX. Furthermore, these two types of responses may exist in a myotube at the same time. Under some circumstances the kinetics of the two systems are sufficiently different to result in action potentials that have two peaks. Under these conditions the first peak is always of shorter duration and it is always blocked by TTX.
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  • 125
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: Measurements of simultaneous mitotic activity, electrical transmembrane potential (Em), and cell density levels in both 3T3 and Chinese hamster ovary (CHO) cell cultures reveal that a 5- to 6-fold increase in the Em level is associated with development of mitotic arrest at saturation densities. This rise occurs both in confluent monolayers and in interior areas of isolated colonies, and is independent of the rate at which confluence is attained. The Em rise is accompanied by a substantial decrease in intracellular Na. Electron microscopy of saturated CHO monolayer sections shows from 46 to 63% of the cell surfaces to be in close apposition (〈300 Å spacing). These results for contact inhibited cultures support the hypothesis that mitotic activity may be functionally coupled with the Em level and associated ionic concentration levels. It is suggested that contact inhibition of mitosis may result from a reduction in synthesis of mitogenically essential RNA following a decrease in intracellular Na produced by contact-induced alteration of surface ion-transport activity.
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  • 126
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The mechanically stimulable bioluminescence of members of the Gonyaulax catenella group can be maximally photoinhibited by exposure to as few as 1013 quanta/cm2, a factor 104 times smaller than that required for comparable photoinhibition in Gonyaulax polyedra and all other photosynthetic bioluminescent dinoflagellates investigated. Following an irradiation pulse there is an initial time lag of one minute, followed by a rapid decrease in mechanical stimulability to approximately 1% of the dark unirradiated control with a firstorder rate constant as high as 0.01 sec-1. Action spectra for all three species imply a pigment with a single absorption band having a maximum at 562 nm and a half band width of 105 nm within the spectral range 325 nm to 775 nm. Photoinhibition appears to decrease either the sensitivity of the shear receptor mechanism or the efficiency of signal transmission in the dinoflagellates, since chemically stimulable bioluminescence is unaffected by these exposures.
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  • 127
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of colchicine and 2-Br-α-ergocryptine-methane-sulfonate (CB 154) on the release of prolactin and growth hormone have been studied in a clonal strain of rat pituitary tumor cells (GH3) in monolayer culture. These cultures produce both prolactin and growth hormone and release both proteins spontaneously into the medium without storing them in large amounts. Immunological methods were used to measure both intracellular and extracellular concentrations of the hormones. Colchicine (5 × 10-6 M for 3 hours) caused a 2- to 3-fold increase in intracellular concentrations of prolactin and growth hormone but, under basal conditions, had little or no measurable effect on the amounts of hormone accumulated in the medium during the course of the standard three hour treatment period. This latter finding evidently is due to a lag in the onset of drug action. Colchicine had little or no effect on accumulation of extracellular prolactin during the first two hours of treatment whereas such accumulation was depressed by over 60% during the third hour of treatment. Previous studies have shown that treatment of GH3 cells with thyrotropin releasing hormone (TRH) and hydrocortisone (HC) increases both intra and extracellular levels of prolactin and growth hormone, respectively. In cultures treated with TRH (5 × 10-8 M), colchicine (5 × 10-6 M for 3 hours) increased intracellular prolactin by about 70% and decreased extracellular hormone by 10%. In cultures treated with HC (3 × 1O-6 M), colchicine increased intracellular growth hormone by more than 100% and decreased medium concentrations of the hormone by 15%. Colchicine did not significantly alter total hormone (intracellular + extracellular) accumulation, cellular uptake of 3H-amino acids, or total cell protein synthesis. The synthetic ergot alkaloid, CB 154, (3.3 × 10-6 M for 3 hours) caused an 80% increase in intracellular, and a nearly 50% decrease in extracellular, prolactin without affecting the accumulation of growth hormone, the uptake of 3H-labeled amino acids, or overall protein synthesis in the cultures. Elevation of medium potassium concentration from a basal value of 5.3 mM to 3-5 × 10-2 M (by addition of KCl) decreased intracellular levels of prolactin by 85% and growth hormone by 55%. These effects of high potassium were blocked by colchicine and by CB 154. We conclude that colchicine, after a lag period of two hours, acts to inhibit the release of prolactin and growth hormone from GH3 cells. By the end of three hours of treatment, this inhibition is over 60% complete in the case of prolactin. The qualitatively different effects of colchicine and CB 154 on prolactin and growth hormone release suggest that these two secretory blocking agents probably act on GH3 cells by different mechanisms.
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  • 128
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    Journal of Cellular Physiology 82 (1973), S. 435-444 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have investigated the effects of the amino reactive reagent, 2,4,6-trinitrobenzene sulfonic acid (TNBS) on anion transport (chloride and sulfate) and on the K+ content of Ehrlich ascites tumor cells. Incubation of tumor cells with TNBS (3 mM or 10 mM) results in a time dependent uptake of this molecule. Tightly bound TNBS caused a loss of K+ as well as inhibition of sulfate uptake. Although sulfate transport was inhibited by tightly bound TNBS (40% inhibition with 20 nmoles bound per 107 cells), reversibly bound TNBS exerted much greater inhibition. Kinetic analysis of sulfate transport in the presence and absence of TNBS suggests that: (1) tightly bound TNBS exerts a competitive inhibition by occupying membrane sites remote from the specific transport site, (2) TNBS reversibly interacts with a separate site also in a competitive fashion.Increasing amounts of tightly bound TNBS resulted in an enhanced chloride influx. However, reversibly bound TNBS was without effect. These results are in contrast to the effect of TNBS on sulfate transport and show that TNBS, at least in this cell type, is not a general inhibitor of anion transport.
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  • 129
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    Journal of Cellular Physiology 82 (1973), S. 513-513 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 130
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    Journal of Cellular Physiology 81 (1973), S. 355-363 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cell proliferation in density-inhibited chick embryo cell cultures was induced by microgram quantities of insulin, neuraminidase, trypsin or papain. Other proteins tested, including albumin, fetuin, ribonuclease and hyaluronidase were inactive except in very high concentrations (〉 100 μg/ml).The insulin chick embryo model was selected for detailed analysis of the initiation of proliferation. Insulin insolubilized by conjugation with Sepharose particles was also active, but only in so far as it was released in soluble form from the particles. This was measured by a radioimmunoassay. Under the conditions giving maximal cell proliferation less than 0.002-0.2% of insulin was taken up by the cells. This suggests that an interaction of insulin with the cell surface only is sufficient to stimulate the cells. Insulin released the density-inhibited cells from G1 phase to produce an almost synchronous wave of proliferation. The following sequence of events was characteristic of the cells after stimulation by insulin: an early increase in sugar uptake and decrease in leucine uptake, increase in cell volume, stimulation of RNA and protein synthesis, increase in thymidine uptake, DNA synthesis, mitosis and cell division.
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  • 131
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: The differentiated state of mouse erythropoietic progenitor cells (CFU-E), detected by their ability to form erythropoietin-dependent colonies in vitro, has been investigated. Transfusion-induced plethora was found to reduce the population size of CFU-E in both spleen and femoral marrow, which indicates that a significant number of CFU-E arise by differentiation processes that are themselves erythropoietin-dependent. Individual spleen colonies were found to be heterogeneous in their content of CFU-E, and the numbers of CFU-E per colony were not correlated either positively or negatively with the numbers of granulocyte-macrophage progenitors (CFU-C) present in the same colonies. The absence of a negative correlation between CFU-E and CFU-C indicates that the erythropoietic and granulopoietic pathways of differentiation are not mutually exclusive within individual spleen colonies. The numbers of CFU-E per spleen colony were also found to vary independently of the numbers of pluripotent stem cells (CFU-S) per colony; in contrast, as found previously, the numbers of CFU-C and CFU-S per colony were positively correlated. These results indicate that more randomizing events separate CFU-E from CFU-S than separate CFU-C from CFU-S, and are consistent with the view that CFU-E occupy a position on the erythropoietic pathway of differentiation that is more remote from the pluripotent stem cells than is the corresponding position of CFU-C on the granulopoietic pathway.
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  • 132
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    Topics: Biology , Medicine
    Notes: Calcium uptake by normal human lymphocytes was found to be a saturable process which was competitively inhibited by manganese indicating the existence of a carrier-mediated mechanism for calcium uptake. Exchange diffusion was not observed, Phytohemagglutinin (PHA) significantly stimulated calcium uptake within minutes after treatment. The increased uptake was attributed to a decreased Km for the proposed membrane carrier rather than to an increased Vmax. Also PHA did not stimulate a normally unused exchange diffusion process, nor did it affect calcium efflux. Uptake by both unstimulated and PHA-treated lymphocytes was not influenced by magnesium or by cycloheximide or actinomycin D.
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  • 133
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    Journal of Cellular Physiology 82 (1973), S. 65-73 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Purified subcellular fractions containing melanosomes from B-16 mouse melanoma were treated with 2% sodium dodecyl sulfate or 0.5 M sodium hydroxide to dissolve protein. Quantitative measurements indicate that each melanosome contains 0.065×10-10 of protein or about 19% by weight. SDS acrylamide gel electrophoresis of proteins from purified melanosomes resolved six polypeptide bands of major density and about 15 minor bands. These results indicate that the melanosome may be more complex than previous genetic, biochemical or morphological evidence had suggested.
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  • 134
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: 3T3 mouse fibroblasts in 10% calf serum cease to increase in number at confluence. SV40-transformed 3T3 cells lose their sensitivity to this growth control and so are able to reach multilayered cell densities. We previously described a polyploid revertant line F1SV101, isolated from a SV40-transformed cell, that has a low maximum cell density (Pollack et al., '68), despite the continued presence in this revertant of SV40-specific DNA, RNA and Tantigen. We have examined the mechanism by which the revertant maintains a low saturation density, and find that at confluence the revertant and 3T3 are both reduced in mitotic index, fraction of cells synthesizing DNA, and rate of DNA synthesis. The transformed cell does not respond to confluence. None of the lines sheds intact cells into the medium at confluence.
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  • 135
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    Notes: The cyclic-AMP concentration in the liver remnant after 70% hepatectomy increases in a biphasic manner with peak values at 3 and 12 hours, and DNA synthesis begins at 18 hours. Propranolol (dl) injected at 30 minutes after surgery stopped the first wave of cyclic-AMP accumulation, but did not affect the second accumulation or the initiation of DNA synthesis. However, dl, propranolol injected at eight hours equally delayed (by 6 to 8 hours) the second wave of cyclic-AMP accumulation and the initiation of DNA synthesis. Propranolol (dl) did not affect DNA replication per se, since it was totally ineffective after the second wave of cyclic-AMP accumulation had passed and DNA synthesis had been initiated. Propranolol (dl) action was not due to a blockade of β-adrenergic receptors, since its d or l isomers were separately without effect, as were unrelated β-adrenergic blockers (Kü 1313 and M&B 17-803A).On the other hand, an activation of α-adrenergic receptors may be involved in the induction of hepatocyte proliferation, since α-adrenergic antagonists, such as phenoxybenzamine and phentolamine, both delayed and considerably reduced the second wave of cyclic-AMP accumulation and the subsequent initiation of DNA synthesis. It is concluded that the second wave of cyclic-AMP accumulation is somehow associated with the initiation of DNA synthesis.
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  • 136
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    Journal of Cellular Physiology 82 (1973), S. 213-218 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The temperature dependence of the uptake of glucose by exchange transport is investigated at two pH values (7.5 and 5). The Arrhenius's energy of activation, the heat of activation, the free energy of activation and the entropy of activation are calculated. The parameters are different at pH 7.5 and 5. For both pH levels the heat of activation and the entropy of activation change abruptly at 20°C. This finding leads us to assume that at this temperature a change in the structure of the membrane takes place.
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  • 137
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Previous studies showed that after 5 μg of Salmonella typhosa endotoxin there was an increase in colony stimulating factor temporally related to a fall in murine marrow in vitro colony forming cells (CFC). This was followed by differentiation along the marrow granulocytic pathway. The present studies showed that after 5 μg of endotoxin the peripheral blood CFC fell by approximately 50% at one hour, rose to a level ten fold that of control at six hours and then returned to control values by 48 hours. There was a progressive increase in the number of splenic CFC to ten fold that of control from 24 to 72 hours after endotoxin. These data imply a migration of CFC from the marrow to the spleen along with an in-situ increase in splenic CFC. Thus, either migration or differentiation may explain the fall in marrow CFC after endotoxin.Spleen colony forming units (CFU) in the marrow were measured by a transplantation technique and the transplantation fraction (f Fx) determined. A decrease in marrow CFU at 24 hours after endotoxin was secondary to a change in the f Fx. from 11.1% to 7.6%. There was however, an increased percentage of CFU in DNA synthesis in the interval of 6-48 hours after endotoxin, as judged by the hydroxyurea technique. As the marrow CFC fell within 20 minutes of endotoxin administration, the data suggest the CFC may be affected initially and that changes in the generative cycle of the CFU may be of a secondary nature.
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  • 138
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    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Medium conditioned by excised whole lungs from endotoxin-injected C57BL mice was highly active in stimulating hemopoietic colony formation, particularly of granulocytic type, in agar cultures of mouse bone marrow cells. The colony stimulating factor (CSF) in this material had an α1-α2 electrophoretic mobility, was eluted from calcium phosphate gel by 0.04 M phosphate buffer and had an unusually low apparent S20W of 1.9. Sequestered polymor-phonuclear neutrophils were excluded as a major source of this CSF. The high specific activity and ease of preparation of lung conditioned medium make it valuable both for the large scale production of CSF and as a source of an unusual type of CSF.
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  • 139
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    Journal of Cellular Physiology 81 (1973), S. 55-61 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Subpopulations of human lymphoid cells are capable of spontaneously fusing with fibroblasts of human or murine origin to form human-human or human-mouse hybrid cells. These cells were present in thymus, spleen, and bone marrow. After fractionation on discontinuous bovine serum albumin gradients, the cells were found in the less dense layers of the gradient. Cells of fetal origin, except for bone marrow, fused spontaneously at a higher rate than those of adult origin. The highest rate of fusion was found with adult bone marrow cells. These lymphoid cells appear to be “thymus-derived” cells.
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  • 140
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    Journal of Cellular Physiology 81 (1973), S. 241-250 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Calcium is a major regulator of thymic lymphoblast proliferation in vivo and in vitro. The proliferative activity of the lymphoblasts in thymic lymphocyte (thymocyte) populations in vitro is both constant and low in the presence of calcium concentrations between 0 and 1.0 mM, but higher concentrations increase proliferation by an endogenous cyclic AMP-mediated promotion of the initiation of DNA synthesis.Lower concentrations (10-7 to 10-5 M) of exogenous cyclic AMP (but not 5′-AMP) stimulate lymphoblast proliferation in a low-calcium (0.5 mM) medium, but higher concentrations do not. However, all exogenous cyclic AMP concentrations between 10-7 and 10-3 M (but again not 5′-AMP) block the stimulation of lymphoblast proliferation in a high-calcium (1.5 mM) medium. Exogenous cyclic AMP does not prevent calcium from “activating” lymphoblasts, but it reversibly blocks the reaction responsible for the initiation of DNA synthesis in these stimulated cells. Finally, cyclic AMP's inhibitory action, in contrast to its stimulatory action in low-calcium medium, is not specific for the cyclic nucleotide since a low, non-mitogenic concentration of cyclic GMP also prevents calcium from stimulating DNA synthesis and cell proliferation.
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  • 141
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: The induction, isolation and some of the properties of serine-requiring mutants of Chinese hamster ovary cells have been described. These cells promise to be useful in biochemical genetic studies and cancer research.
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  • 142
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    Journal of Cellular Physiology 81 (1973), S. 339-345 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Purine nucleotides and nucleosides were found to enhance the DNA synthesis in quiescent 3T3 cells induced either by fresh medium containing calf serum or by the addition of the β-globulin serum fraction to depleted medium. A two to threefold enhancement has been obtained under these conditions. The adenine and hypoxanthine based compounds were more effective than the guanine ones in the concentration range tested (10-5--10-3 M), while the pyrimidine compounds were without effect. The degree of enhancement obtained with the purine compounds depended on the type and amount of serum used to stimulate DNA synthesis. A maximal degree of enhancement was obtained when the purine compounds were present for only the first three hours after the addition of serum and thus 18-20 hours before the actual onset of DNA synthesis.
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  • 143
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    Journal of Cellular Physiology 81 (1973), S. 397-410 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Clonal growth in semisolid agar medium was obtained using cells from 19 of 25 transplanted murine plasmacytomas when the medium was supplemented by whole mouse blood or washed red cells. With different tumors cloning efficiency ranged from 0.01% to 21.6%. With two exceptions, mouse blood did not potentiate colony formation in agar by cells from transplantable myelomonocytic, myeloid, and lymphoid leukemias, reticulum cell sarcomas and fibrosarcomas. The clonal growth of some plasmacytomas was also potentiated by syngeneic thymic, spleen or bone marrow cells. Plasmacytoma colony growth was not stimulated by normal mouse serum but serum from mice injected with endotoxin or polymerised flagellin stimulated colony growth by some plasmacytomas. The active serum factor was not the colony stimulating factor (CSF) and its appearance after antigenic stimulation was not T cell-dependent. Preimmunised mice failed tq respond to antigenic stimulation. Whole body irradiation did not induce a rise in the capacity of serum to stimulate colony formation by plasmacytoma cells.
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  • 144
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    Journal of Cellular Physiology 81 (1973), S. 387-396 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Chick embryo cultures deprived of serum synthesize DNA at a reduced rate. DNA synthesis in serum-deprived cultures is stimulated as much as ten-fold by the addition of Zn++, Mn++ or Cd++ in concentrations just below the toxic level. These metals, in the same concentration range, also stimulate the uptake of 3H-2-deoxy-D-glucose (2-DOG). The increase in uptake of 2-DOG precedes the increase in synthesis of DNA, and is probably an indicator of a more general membrane perturbation. The metals also stimulate DNA synthesis in serum-containing, density-inhibited cultures. The carcinogenic hydrocarbon 9,10-dimethyl-1,2-benzanthracene stimulates DNA synthesis and 2-DOG uptake in serum-deprived cultures at those concentrations which also cause morphological changes in the culture. Other carcinogenic hydrocarbons, which produce no morphological changes in the culture do not stimulate DNA synthesis. In contrast to these non-specific effects, DNA synthesis which is inhibited by low concentrations of either ethylene diamine tetraacetate (EDTA) or diethylene triamine pentaacetate (DTPA) is stimulated specifically by Zn++. These findings are interpreted to mean that certain metals and carcinogens, like a variety of other agents, interact non-specifically with the plasma membrane to initiate a chain of events leading to DNA synthesis, and that one of these events is the liberation of Zn++ for enzyme reactions leading to DNA synthesis.
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  • 145
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    Topics: Biology , Medicine
    Notes: By mapping the location of isolated single cells; and then counting the number of cells at each location as a function of time. it was possible to accumulate data on the growth history for each of a large group of clones. The clone size distribution, its mean and standard deviation were computed for each day in culture. Variations in schedule of medium change and time of exposure to trypsin, did not measurably affect variation in clone size. Neither could clone size variation be accounted for on the basis of (1) occurrence of nondividing cells nor (2) presence of heritable growth rate variants in the population. It is probable that clone size variation under our conditions is primarily a consequence of a highly variable interdivision time among the constituent cells.
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  • 146
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    Journal of Cellular Physiology 82 (1973), S. 113-120 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The synthesis of immunoglobulin and of nuclear proteins has been studied in synchronized mouse myeloma cells of the C1 line. Synchronization has been obtained by a double thymidine block. C1 cells synthesize immunoglobulin at a relatively constant rate throughout the cell cycle except for mitosis, when a decrease in the rate of synthesis of total protein and of immunoglobulin is observed. Cell synchrony around mitosis is not sufficiently good to determine whether immunoglobulin is synthesized at all. Nuclear protein and in particular histones appear to be synthesized synchronously with DNA during the S phase of the cell cycle.
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  • 147
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    Journal of Cellular Physiology 82 (1973), S. 165-179 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The biosynthesis of pyridine nucleotides has been examined in a number of mammalian cell lines in culture. In all lines examined, nicotinamide is incorporated by a biochemical pathway distinct from the Preiss-Handler pathway for nicotinic acid.In at least the human cell line D98/AH2, there is no detectable endogenous synthesis of the pyridine ring from tryptophan. Although most cell lines examined (hamster BHK 21/13, mouse L929 and human D98/AH2) use either nicotinic acid or nicotinamide as a precursor for DPN and TPN, two mouse cell lines, 3T3-4E and LM CIID, are unable to utilize nicotinic acid as a source of the pyridine ring.If nicotinic acid is present in the medium, substantial amounts of intracellular desamido DPN accumulate suggesting that the last step (desamido DPN→DPN) is limiting in the Preiss-Handler pathway. With nicotinamide, the only compound which accumulates in substantial amounts apart from DPN and TPN is nicotinamide ribose; there is no detectable NMN. The results of pulse-labeling experiments suggest that nicotinamide ribose may be an intermediate in the nicotinamide pathway.Following growth of D98/AH2 cells in high concentrations of niacin, biosynthesis of DPN from nicotinamide was completely inhibited for at least six hours. The converse experiment revealed no inhibition of niacin incorporation. This observation suggests that a niacin pathway intermediate, which present evidence indicates is desamido-DPN. can inhibit nicotinamide utilization.Newly synthesized DPN turns over with a half-life of two hours in azaserine-treated D98/AH2 cells. In the absence of azaserine, the nicotinamide moiety of newly synthesized DPN is lost from D98/AH2 cells to the medium with a half-life of eight hours. About 80% of the nicotinamide is lost to medium as nicotinamide ribose.
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  • 148
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    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The average speed of nuclear translocation of 3T3 cells, recorded in a time-lapse film of a perfused culture, was negatively correlated with the number of contacting cells, and, to a lesser degree, with the amount of a cell's perimeter in contact with other cells. When a cell was in contact with five or more other cells, its speed was reduced by 50%, on the average, although the variation in individual cell speed was considerable at each level of contact. A partial correlation analysis showed that any extracellular soluble factors governed by the local cell density had little or no effect on speed, relative to the prominent effect of the number of cell-cell contacts, and hence that 3T3 cells display true contact inhibition of speed. This confirms the original demonstration by Abercrombie and Heaysman (1952), who studied chick embryo heart fibrpolasts. In our study, the relationships between average speed and age of the culture was such that a possible independent contribution of a time-associated factor other than contact to the diminution in average speed, although not necessary to account for the data, could not be excluded.The same intercellular contacts found to inhibit speed in this study were previously reported to cause no immediate prolongation of individual cell generation times, despite the fact that the filmed culture was undergoing so-called “contact” inhibition of cell division. In the present study, moreover, no correlation was observed between the average speeds of individual cells and their generation times. Hence, postconfluence inhibition of cell division and contact inhibition of speed of cell movement seem to be independent phenomena.
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  • 149
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: Erythroid differentiation of Friend leukemia cells is enhanced when the cells are grown for four days in the presence of dimethylsulfoxide (DMSO). Dimethylformamide (DMF) has a similar though less marked effect. 5-Bromo-2′-deoxyuridine (BUdR) (10-5M) inhibits both DMF- and DMSO-stimulated differentiation. For maximum inhibition, BUdR must be present during the first two days of growth, during which time DNA synthesis is maximal. The addition of BUdR after the third day has no effect.Since BUdR is incorporated into DNA and thymidine prevents BUdR inhibition of DMSO-stimulated differentiation, it is likely that BUdR acts by virtue of its incorporation into DNA. Although BUdR alone had little effect upon cell multiplication, in combination with DMSO, cell growth was inhibited up to 40%. Since the BUdR-inhibition of the DMSO effect was approximately 70%, it is unlikely that its effect on differentiation is due to selective killing of those cells which are stimulated to differentiate.
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  • 150
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: Flow microfluorometry has been used to quantitate cell-surface binding of fluorescein-conjugated lectins. Frequency distributions of total surface binding of Concanavalin A per cell were prepared for a variety of cultured cell populations, including established cell lines, virus-transformed lines and non-transformed parental lines. In the case of growing Chinese hamster cells (line CHO), much of the variability of Con A binding per cell could be related to variability of cell size. Experiments with cells synchronized by mitotic selection indicated that the modal surface density of binding sites was almost constant throughout the cell cycle. However, as indicated by inhibition of binding with α-methyl mannopyranoside and by the effect of trypsin, the sites on each cell were heterogeneous in chemical structure and/or exposure. Agglutinability of virus-transformed cell lines or trypsin-treated parental lines was demonstrated but could not be correlated closely with binding.
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  • 151
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    Journal of Cellular Physiology 81 (1973), S. 315-321 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: An adenosine kinaseless (AK-) mutant of the mouse fibroblast line 3T6 has been obtained in cell culture by evolution of resistance to 6-thio-methylpurine ribonucleoside and tubercidin. The mutant excretes purines (xanthine and hypoxanthine) into the culture medium. Human or mouse cells lacking hypoxanthine-guanine phosphoribosyl transferase (HPT-) excrete increased amounts of purines, but a human cell mutant lacking both HPT and AK excretes considerably more hypoxanthine. The difference in hypoxanthine excretion between the HPT- mutant and the HPT- AK- mutant originates from the adenosine normally reutilized through the activity of adenosine kinase. The activity of adenosine kinase is essential to retard the adenosine cycle and to prevent cellular loss of purines.
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  • 152
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Leukemic myeloblasts and cells derived from normal chick hematopoietic tissue produced colonies in soft agar. Colonies produced by leukemic myeloblasts differed from normal chick tissue in their morphological characteristics, in the greater initial number of cells required for colony formation and in their decreased dependence on conditioned medium for development. The colony forming cells for both types were enriched when allowed to grow for several days in liquid growth medium.In soft agar, myeloblasts differentiated into more mature granulocytic cells and macrophages. These differentiated cells accumulated between one and two weeks after seeding. When tested for release of avian myeloblastosis virus (AMV), 6 out of 18 colonies were releasing AMV at one week whereas 3 out of 39 were releasing AMV at two weeks. Five two week old colonies which were negative for AMV were producing myeloblastosis associated viruses (MAVs). Normal colony forming cells were present in leukemic buffy coat and although colonies made by these cells contained MAVs, no AMV could be detected.The data obtained with normal avian tissues were similar to those obtained by others with mammalian hematopoietic tissue. Colony formation by normal hematopoietic tissues was strictly dependent on factors present in conditioned medium. Tissues producing colonies included bone marrow, yolk sac, spleen and peripheral leukocytes. Colonies were not obtained from thymus and bursa. Furthermore, the colony origin did not appear to be erythroid in nature.
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  • 153
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    Journal of Cellular Physiology 82 (1973), S. 1-8 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Data have been presented with respect to the widely varying pH optima for the growth of a number of normal and transformed human, mouse, monkey, hamster, rabbit and rat cell lines. At that optimum, differences between normal and transformed cells with respect to maximum population densities tended to become less prominent. Other differences (serum requirement, long-term viability of cultures) were not demonstrably pH-dependent.
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  • 154
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Granulocyte-macrophage colony formation from bone marrow cells in soft agar is dependent upon the presence of a stimulating factor and the number of colonies is related to its concentration. This dose-response effect provided a measurement of the responsiveness to stimulation of colony forming cell populations in marrows from different sources. There were significant differences between the responsiveness of cells from different strains of mice which paralleled the previously observed myelopoietic and immune responsiveness of these strains to stimulation in vivo.Low concentrations of hydrocrotisone reduced the responsiveness of colony forming cells (a) when added to cultures of normal marrow or (b) when cells were taken from hydrocortisone-treated mice and cultured in its absence. The reduction which followed inoculation was not apparent until the 4th day and occurred irrespective of mouse strain, type of drug or route of inoculation and with a dose (100 μg) which did not affect the actual number of colony forming cells in the marrow.
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  • 155
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Sedimentation velocity separation of Rhesus monkey bone marrow cells has demonstrated a reproducible but heterogeneous size distribution of cells capable of forming granulocytic colonies in agar culture (CFC's). This heterogeneity is shown to be due to the cell cycle status of the progenitor cell population. In vitro exposure of bone marrow cells to lethal doses of tritiated thymidine (H3TdR) either before or after separation restricts the size distribution of CFC's, greatly reducing the proportion of rapidly sedimenting cells. The calculation of the volume distribution of such cells before and after H3TdR exposure indicates that 55% of total CFC's in adult marrow are in G0 or G1 with a volume of 410 μ3, 42% are in S phase and of volume 450-950 μ3, and the remainder are in G2 and mitosis with a volume of between 600-950 μ3. CFC's in mid gestation fetal liver were larger than their adult counterparts and were of homogeneous volume indicative of a single non cycling population with no evidence of an S or G2 component. H3TdR exposure confirmed the non-cycling status of these fetal progenitor cells.
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  • 156
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    Journal of Cellular Physiology 82 (1973) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 157
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    Journal of Cellular Physiology 82 (1973), S. 129-131 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: If the aging of diploid human fibroblasts reflects stable genetic or epigenetic changes which are few in number and different in different cells, then complementation could occur in hybrids between individual senescent cells. However neither pairs of aged fibroblasts nor even pairs of young and senescent fibroblasts produce viable hybrids under conditions known to promote cell fusion.
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  • 158
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    Notes: We have shown that the murine lymphoblast, L5178Y, requires extracellular magnesium or calcium for proliferation in suspension culture. Although both cations produce biphasic effects on growth, magnesium is the more potent since it: (1) stimulates proliferation at lower concentrations; (2) supports optimal proliferation over a wider concentration range; (3) maintains higher cell densities at stationary phase; and (4) produces less inhibition at high concentration.At suboptimal concentrations, calcium facilitates the effect of magnesium but at optimal concentration, no facilitation is evident. A concentration of 3.2 mM calcium or magnesium inhibits growth, while the same concentration composed of equimolar calcium and magnesium does not inhibit proliferation.Extracellular calcium and magnesium may influence proliferation by effecting changes in intracellular calcium and magnesium since: (1) reduction in extracellular concentrations sufficient to produce decreased growth rate is associated with decreased cell calcium and magnesium; (2) cell magnesium and growth rate are related such that an exponential decrease in relative doubling time occurs with decrease in cellular magnesium.Cells cultured at 37°C in magnesium and calcium-deprived medium will proliferate at a normal rate if either or both cations are replaced within six to 10 hours; however, with longer deprivation the doubling time in restored medium is progressively lengthened.
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  • 159
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: Fluoride, iodoacetate, oxamate, 2,4-dinitrophenol, cycloheximide, and ouabain were studied to determine if any of these inhibitors affected the intracellular concentration of sodium and potassium in an L cell strain of mouse fibroblasts and to determine if the changes observed in these parameters could be correlated with growth rate. The results indicated that (1) the intracellular concentration of sodium and potassium could not be correlated with growth rate, (2) fluoride, iodoacetate, oxamate, 2,4-dinitrophenol, and cycloheximide at concentrations having an equal effect on growth had a similar effect on the intracellular sodium and potassium concentration These changes were not as great as those seen with ouabain, which at a concentration which did not inhibit growth, had an equal or greater effect on the intracellular sodium and potassium concentration.
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  • 160
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    Journal of Cellular Physiology 82 (1973), S. 121-128 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The mode of action of the antibiotic, trichodermin, on yeast cells has been investigated. Trichodermin specifically inhibits protein synthesis and, during the in vivo inhibition of protein synthesis, ribosomes remain in polyribosomes rather than shifting to monoribosomes. This observation suggests that trichodermin inhibits either an elongation step or a termination step of protein biosynthesis. These two possibilities were distinguished by comparing the action of trichodermin with that of cycloheximide, a known elongation inhibitor, upon the reformation of polyribosomes during recovery from a block in polypeptide chain initiation. Cycloheximide slows the recovery of polyribosomes from monoribosomes following a block in polypeptide chain initiation whereas trichodermin enhances the recovery of polyribosomes. This observation is interpreted to mean that trichodermin primarily inhibits the termination step of protein biosynthesis.
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  • 161
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    Journal of Cellular Physiology 82 (1973), S. 151-156 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: PMA (phorbol myristate acetate, i.e., 12-0-tetradecanoyl-phorbol-13-acetate) a tumor-promoting ester from croton oil, at its most effective concentration of 0.05 μg per milliliter, rapidly (within one hour) induces a large fraction of the lymphoblasts in suspended thymic lymphocyte populations to start making DNA, and these stimulated cells later progress into mitosis. This stimulatory PMA action is probably mediated by calcium because it disappears when calcium is omitted from the medium, and PMA strikingly increases the sensitivity of the lymphoblasts to calcium's stimulatory action.
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  • 162
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    Journal of Cellular Physiology 82 (1973), S. 181-188 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Treatment of the SV40 transformed 3T3 cell line SV101 with colchicine permits the isolation of polyploid revertant sublines Which have lower saturation densities than SV101. These low saturation density lines have also reverted to a high serum requirement for growth, and are unable to form colonies in methocel. Normal SV40 has been recovered from these revertants.3T3 cells are more resistant to colchicine than SV3T3 cells at all cell densities. Colchicine revertants do not display a 3T3-like resistance to colchicine at low density, but do survive colchicine at confluent cell densities, presumably due to their increased contact inhibition.
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  • 163
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    Journal of Cellular Physiology 82 (1973), S. 219-230 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Microspectrophotometric absorption measurements were used to determine the hemoglobin content of erythroid cells derived from the yolk sac during gestation of fetal C3H mice, from day 9 to day 15. Using the DNA content as a marker for the mitotic state between 2C and 4C phase, five successive cell generations and their mean hemoglobin contents were distinguished: 12 pg (pg, picogram = 10-12 gm). 22.2 pg, 37 pg, 50 pg and 56 pg. In the final state, nucleated erythrocytes contained 98 ± 22 pg hemoglobin.Erythroid cells derived from the liver were measured on day 15 of fetal gestation. The hemoglobin content of proerythroblasts was below 0.3 pg. The two cell generations in the basophilic state had 0.6 pg and 1.7 pg respectively. Polychromatic erythroblasts yielded a hemoglobin content of 5.1 pg in the first cell generation and 7.5 pg in the second one. Orthochromatic erythroblasts contained 8 pg, reticulocytes 12 pg and mature erythrocytes 28 ± 7 pg hemoglobin.Calculations based on these data suggest that the rate of total hemoglobin synthesis is similar in both yolk sac and liver erythropoiesis. The difference between the final hemoglobin content in nucleated erythrocytes of yolk sac origin and that in hepatic erythrocytes can be explained by the different cell generation times.
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  • 164
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    Journal of Cellular Physiology 82 (1973), S. 267-275 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The rate of 5-3H uridine uptake into Chinese hamster V79 cells and the rate of its incorporation into RNA increase tenfold during the cell cycle. Both reactions exhibit the same apparent Km(1.7 × 10-5 M). Chromatography of acid-soluble material from cells incubated with 5-3H uridine (0.25 μM) at different times of the cell cycle revealed that intracellular uridine was rapidly phosphorylated at all times, even though cells in late S and G2 take up roughly ten times as much uridine as cells in G1. Uridine kinase activity in synchronized cells increases about two and one-half-fold during the same time period, and in exponentially growing cells is not saturated until the external uridine concentration is raised above 200 μM. It is concluded that the change in uridine kinase activity during the cell cycle is not responsible for the tenfold increase in the rate of uridine transport, and that these two processes are independently regulated.
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  • 165
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    Journal of Cellular Physiology 82 (1973), S. 299-307 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The frequency of mutations induced by ethyl methane sulfonate was compared in a pseudodiploid Chinese hamster cell strain and in a tetraploid substrain derived from it. The frequency of reverse mutations from glycine auxotrophy to glycine independence was similar in the two strains, as expected for a dominant phenotype. Forward mutation to 6-thioguanine-resistance was 25 fold lower in the tetraploid as compared to the diploid strain. The resistant mutants lack hypoxanthine phosphoribosyl transferase activity and their resistant phenotype is recessive in somatic cell hybrids. A combination of chromosomal segregation and mutation could account for the frequency of these recessive drug-resistant mutants in the tetraploid population.
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  • 166
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    Notes: Three assays for bone marrow progenitor cells have been used to determine the effect of single doses of two cytotoxic agents, cyclophosphamide and vinblastine. The assays employed were the agar colony forming and spleen colony forming assays and the crythroid repopulating ability.In normal mice, there was little difference between the response of the progenitor cells assayed by the three methods, following cyclophosphamide: and no detectable difference following vinblastine.Bone marrow from continuously irradiated mice and bone marrow regenerating seven days following transplantation was also studied: in both these situations the proliferation rate of the progenitor cells is increased. Cyclophosphamide was found to be only slightly proliferation dependent with each assay. However, vinblastine was strikingly proliferation dependent. In irradiated mice and also in regenerating marrow the agar colony forming cells were many times more sensitive to this agent than were the other progenitor cells.These results show that under some but not all circumstances the agar colony forming and spleen colony forming cells behave similarly in C57BL mice, but are not a single population of cells.
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  • 167
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    Notes: A pseudo-first order kinetic analysis has described the reactions of purine ribonucleotide synthesis and interconversion in Ehrlich ascites tumor cells in vitro. Rate constants were determined for 13 reactions at non-saturating precursor concentration (50 μM); nucleotide formation from adenine-14C and hypoxanthine-14C was rate limiting for ATP synthesis and nucleotide formation from guanine-14C and hypoxanthine-11C was rate limiting for GTP synthesis. The initial rites of nucleotide synthesis from 1.0 mM adenine-14C and hypoxanthine-14C were equivalent to the rates of nucleotide synthesis at non-saturating base concentration, but the rate of nucleotide synthesis at saturating base concentration after 30 minutes incubation was 20-25% that of non-saturating precursor concentration.
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  • 168
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    Notes: Recent studies have suggested that ectosulfhydryl groups may play a role in cell contact phenomena.We have studied the possible role of ecto- and endosulfhydryl groups in the morphology, adhesiveness, random and directed (chemotaxis) motility and phagocytosis of human polymorphonuclear neutrophils. The rapidly penetrating sulfhydryl binding reagents HgCl2 and NEM inhibited adhesiveness, motility and phagocytosis when studied at 〉 0.1 mM in plasma or 〉 0.01 mM in buffer. The difference in inhibitory concentration was shown to be due to the difference in albumin content of the two media. D-cysteine prevented the effect of HgCl2 and NEM on cell morphology, adhesiveness, motility and phagocytosis indicating that their effects were on cell sulfhydryl groups. PCMBS, a very slowly penetrating organic mercurial, had no effect on neutrophil morphology, adhesiveness, motility or phagocytosis. However, PCMBS inhibited platelet aggregation, assuring its potency. These studies indicate that ectosulfhydryl groups are either not present or not participants in the maintainence of structure and functions of human neutrophilic granulocytes.
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  • 169
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    Journal of Cellular Physiology 82 (1973), S. 461-473 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A liquid culture system, for haemopoietic cells, has been developed using bone marrow cells alone, or co-cultures of thymus and bone marrow cells, inoculated into four ounce medical bottles. After several days growth, such cultures consisted of an attaching population of cells, forming discrete colonies, and a non-attaching population. In the (co-cultures) there was a 2 X enhancement of monolayer colony development compared with the combined total present in the (marrow alone) plus (thymus alone) cultures. Also, better maintenance of non-attaching cells was seen in the (co-cultures). Normal CFUS and CFUC were present in both the (marrow alone) and the (co-cultures) for at least 14 days.In the (marrow alone) cultures, granulocytes in all stages of development were present for the first week, but by 12 days the culture consisted mainly of mono-nuclear cells. In the (co-cultures), however, at 12 days more than 60% of the cells were granulocytes, in all stages of differentiation. (Co-cultures) established using lethally irradiated thymus cells were not able to support this prolonged myeloid differentiation.By feeding the (co-cultures) it was possible to maintain production of (granulocytic) cells for at least ten weeks, although no fully mature granulocytes were observed. After the second feeding, no CFUS were detectable, but variable numbers of agar colony forming cells (not classical CFUC) were present at least for ten weeks.
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  • 170
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    Notes: We have found that diploid human fibroblasts, but not heteroploid human fibroblasts, stringently required serum for their multiplication. Using Diaflo ultrafiltration membrane units, isoelectric focusing and preparative gel electrophoresis, we have isolated and purified this mitogenic activity from mammalian sera. This electrophoretically homogeneous sialoprotein is 120,000 daltons in size, with an iso-electric point of pH 5.2-5.4; it is made up of two electrophoretically identical dimers weighing 57,000 each; it is thermostable and is inactivated by both trypsin and neuraminidase.
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  • 171
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  • 172
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    Journal of Cellular Physiology 81 (1973), S. 347-353 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Somatic hybrids obtained by the selective method of Littlefield between a permanent line of Chinese hamster cells (Wg3) and one of mouse cells (3TP) showed a preponderance of biarmed (hamster) chromosomes. Under normal culture conditions (37°) the doubling time of the parental mouse cells was twice as long as that of the parental hamster cells. If the temperature of incubation was lowered (31°), the relative difference in doubling times was reduced; in hybrid lines obtained under these conditions, the proportion of biarmed chromosomes was also reduced.Upon extended cultivation the average number of telocentric chromosomes progressively decreased in all hybrid lines tested, regardless whether these were started and maintained at 37° or at 31°. An inverse correlation was observed in hybrid cells between doubling time and relative proportion of biarmed chromosomes, suggesting that the karyotypic changes observed after extended culture were due to the selective overgrowth of cells with a high biarmed to telocentric ratio.
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  • 173
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    Notes: Previous studies with mammalian cultured cells have shown that volume regulation in hypotonic medium requires active Na transport. In the present study, determinations of intracellular Na and K content were made in cultured mouse lymphoblasts during the process of swelling and subsequent shrinking (volume regulation) in hypotonic medium. Na and K content were measured in cells in which the shrinking phase was inhibited by the cardiac glycoside, ouabain. In osmotically-shocked cells, an initial permeability increase to K, and not Na, was observed, which allowed K to diffuse out rapidly, down its gradient. Na, meanwhile, rapidly flowed inward with water entry during the swelling process, and was later lost with the same kinetics as the cell shrinkage. This loss of Na was prevented in the presence of ouabain. The results imply that volume regulation is achieved by pumping Na gained during swelling out of the cells, while any K taken up by the pump is rapidly lost through a more permeable membrane. The loss of osmotically active Na, presumably with accompanying anions, allows water to passively diffuse down its osmotic gradient, reducing cell volume subsequent to the initial passive swelling, during which K was rapidly lost.
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  • 174
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    Journal of Cellular Physiology 82 (1973), S. 319-321 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Myoglobin synthesis was compared in cell cultures of leg (red) and breast (white) muscle of chick embryos. In leg muscle cultures a rapidly increasing amino acid incorporation into myoglobin begins two days after muscle cell fusion; in breast muscle cultures no comparable increase was observed. This qualitative difference in cultures of the two muscle cell types provides possibilities for the further study of the mechanism of myoglobin synthesis.
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  • 175
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    Journal of Cellular Physiology 82 (1973), S. 323-323 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 176
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    Journal of Cellular Physiology 82 (1973), S. 325-331 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Temperature sensitive cells have been isclated from Syrian and Chinese hamster cells using a method based on selective detachment from a glass substrate. The Syrian hamster isolates occurred at a high frequency (about 1 in 103) and reverted rapidly; polyoma virus transformation conferred on cells the ability to grow, perhaps abnormally, in agar suspension. A slightly modified isolation technique was applied to Chinese hamster cultures and resulted in the isolation of at least one mutant (from a starting population of 5 × 108 cells) with a spontaneous reversion rate of less than one in 6 × 107. Treatment of the mutant with ethyl methane sulphonate induced reversion. It was concluded that selective detachment provided a useful method for the isolation of conditional lethal mutants of mammalian cells.
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  • 177
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: Confluent quiescent monolayers of aneuploid and euploid cells in culture can be stimulated to proliferate by appropriate nutritional changes. In confluent monolayers of WI-38 human diploid fibroblasts the uptake of cycloleucine is increased three hours after these cells are stimulated to proliferate by a change of medium plus 10% serum. No changes in the uptake of cycloleucine are observed in logarithmically-growing WI-38 cells exposed to fresh medium plus 10% serum, or in WI-38 confluent monolayers in which the conditioned medium has been replaced by fresh medium with 0.3% serum (a change that does not cause stimulation of cellular proliferation in WI-38 cells). In 3T6 cells in the stationary phase stimulated to proliferate by nutritional changes, there is a prompt increase in the uptake of cycloleucine, within one hour after stimulation of cell proliferation. Similar results were obtained with stationary 2RA cells which are SV-40 transformed WI-38 fibroblasts. In addition, chromatin template activity which is known to increase in the early stages after stimulation of confluent WI-38 cells, was unchanged in confluent 3T6 or 2RA cells stimulated to proliferate.These results show that at least two of the very early biochemical events occurring in response to stimulation of cell proliferation are different in WI-38 diploid cells and in aneuploid 2RA or 3T6 cells. It is proposed that WI-38 cells in the stationary phase are arrested in the G0 phase of the cell cycle, while 2RA and 3T6 cells are arrested in the G1 phase.
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  • 178
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    Journal of Cellular Physiology 82 (1973), S. 475-488 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: When Ehrlich ascites cells were cultivated in serum-free media their cellular protein synthetic rate declined to a new steady-state level and the cells stopped multiplying. On addition of serum the cellular protein synthetic rate increased to the level before serum starvation and cells resumed multiplication. The activity in serum stimulating protein synthesis was inactivated on incubation with cells. At cell concentrations of the usual culture conditions this inactivation took several hours; at very high cell concentrations it was complete in ten minutes. Serum-starved cells inactivated low serum (2%-6%) media in the same length of time. Studies of inactivation of high serum media demonstrated that cells had a limted capacity to inactivate. Cells grown in 10% serum were unable to inactivate. Inactivation was not due to accumulation in the medium of either low molecular or macromolecular cell products. Inactivation was strongly inhibited at 4° or by treatment of cells with fluoride or cycloheximide (long exposure): less inhibited by treatment with 2-deoxyglucose or glutaraldehyde; and slightly inhibited by treatment with cyanide or cycloheximide (short exposure). Inactivating ability was unaffected by trypsinization. These findings are best explained by the hypothesis that cells take up the serum activity by endocytosis.
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  • 179
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    Journal of Cellular Physiology 81 (1973) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 180
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    Journal of Cellular Physiology 82 (1973), S. 199-212 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Wild-type cultured cells of the frog cell line ICR 2A give rise to 5-bromodeoxyridine (BUdR)-resistant colonies only when the selecting concentration of the drug is 5 × 10-5 M or lower. The progeny of these colonies multiply in 10-4 M BUdR; resistance is correlated with the absence of a thymidine (TdR)-specific transport reaction with a Km in the range of 2-7 × 10-4 M. All of the TdR transport-deficient (TT-) isolates examined (25) had TdR kinase activity (4% to 100% of wild-type). Variants deficient in TdR kinase activity (5% of wild-type) were obtained by exposing TT-cultures to 10-3 M BUdR. The TK - variants multply continuously in 10-3 M BUdR and retain the phenotype after prolonged culture in the absence of the drug. The frequency with which they occur is increased 20 to 50 fold by prior treatment of the culture with ICR 191, an acridine mustard mutagen.In haploid cells, it would be expected that TK- variants would arise in equal numbers from wild-type and TT- cultures if loss TdR kinase occurred independently of loss of the transport reaction. However, wild-type cells give no colonies resistant to 10-3 M BUdR under conditions the give 1 to 50 colonies per million TT- cells. The TT- phenotype seems to be a required intermediate state in the origin of the TK- phenotype. Therefore, the TK- clones described above are unlikely to be products of mutation at a single genetic locus.
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  • 181
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    Notes: Calcium uptake by mitochondria and fragmented sarcoplasmic reticulum (FSR) isolated from frog skeletal muscle was studied. These fractions were characterized by electron microscopy, succinic dehydrogenase assay and by using mitochondrial inhibitors. With high (100 μM) Ca in the medium, the Ca accumulating capacity of the two fractions was similar. Zinc in concentrations of 5-10 μM in the medium had no effect on Ca uptake by either fraction whereas higher concentration of Zn (25 μM) reduced Ca uptake in both fractions. Five micromolar lanthanum lowered Ca uptake by 70% in mitochondria but had no effect on Ca uptake by FSR. With 10 and 25 μM La, Ca uptake by FSR decreased by 12 and 20% respectively. Addition of La (5 μM) to Ca-loaded mitochondria had no effect indicating that La could only interfere with the Ca binding step and was unable to release Ca that was already stored. In the medium that originally contained low (10 μM) Ca FSR was able to reduce the Ca concentration below 0.1 μM. In contrast mitochondria, although possessing an equal capacity for Ca uptake were unable to accumulate Ca from the medium when Ca was lowered to approximately 4 μM. Presence of 5-10 μM La in the low Ca medium had no effect on the total amount of Ca taken up by FSR in two minutes but reduced the rate of Ca uptake significantly. The relation of the effects of Zn and La on the isolated fractions to their reported effects on the contractile response of skeletal muscle is discussed.
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  • 182
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    Journal of Cellular Physiology 81 (1973), S. 281-284 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A substance promoting the growth of mammalian cells in vitro has been isolated from Bacto-peptone (Difco Labs., Detroit) and identified as 6,8-dihydroxypurine. This compound, which is isomeric with xanthine(2,6-dihydroxypurine), enhances the growth of one Chinese hamster and some other cell lines; xanthine itself is inactive. The biological action of the compound is discussed.
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  • 183
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    Journal of Cellular Physiology 81 (1973), S. 285-290 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: An apparatus is described for the continuous monitoring of the growth of suspension cultures of mammalian cells in normal conditions and following the administration of a drug.
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  • 184
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    Notes: Using a serial selection technique in which Chinese hamster cells were treated first with 8-azaguanine and then subsequently with HAT medium it was found that approximately 15% of azaguanine resistant clones were also resistant to HAT. Several such clones were subcultured and found to be stably resistant to azaguanine, in some cases at a higher level than the usual azaguanine resistant mutants which are HAT sensitive. Measurements of hypoxanthine-guanine phosphoribosyl transferase levels were in some cases lower than the parental line but in three of the clonal lines were higher than the parental strain. The fact that azaguanine resistant lines constitute a biochemically heterogeneous population underscores the importance of careful characterization of mammalian cell culture variants.
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  • 185
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    Journal of Cellular Physiology 81 (1973) 
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  • 186
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: The elucidation of the subcellular localization of enzymes by the classical technique of homogenization followed by differential centrifugation is limited in that it is difficult to determine the effect of the severe disruptive procedures on the normal relationship of the enzymes to their subcellular environment. Attempts have been made to study this problem under less severe limitations; one of the approaches used has been the use of pressure on whole muscle tissue to extract the cellular fluids. In this report we introduce the concept of “comparative extraction” for evaluation of results obtained by this procedure. By comparing the efflux of enzymes of similar solubility and similar size and shape, it is possible to determine the minimal amount of the less easily extractable enzyme which cannot be removed due to compartmentation or binding to cellular particulate structures.Using this concept of “comparative extraction,” we show in this report that at least 35% of the lactate dehydrogenase of chicken breast muscle is restricted in its removal. The data do not definitely resolve the problem of whether the restriction is due to compartmentation of the enzyme within subcellular organelles or binding to subcellular structures.
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  • 187
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    Journal of Cellular Physiology 81 (1973), S. 225-232 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Contact-inhibited somatic cell hybrids were formed between two malignant cell lines lacking contact inhibition of growth. One cell line was HTC-AR1, an azaguanine-resistant subline from the rat hepatoma line HTC +; the other line was the BUDR-resistant mouse L-cell subline L-B82. Hybrids were obtained from selective medium and characterized by chromosomal and enzymic analysis. The hybrids lacked the inducible rat enzyme tyrosine aminotransferase.
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  • 188
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    Notes: Trypsinization of alveolar macrophages depresses lysine transport but has no effect on nucleoside transport. The depression of the initial rate of lysine transport is due to a decreased rate of exchange diffusion presumably secondary to lowered intracellular pools of amino acids. It is shown directly that trypsinization accelerates the efflux of amino acids but not certain other metabolites, and reduces the capacity of the cell to concentrate amino acids.
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  • 189
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    Notes: We present evidence that the chemical requirements among all the bioluminescent coelenterates that have been examined are very similar or identical to those already described for Renilla by Cormier and associates. Components required for luminescence in Renilla were also found in a number of bioluminescent coelenterates examined such as Aequorea, Obelia, Cavernularia, Ptilosarcus, Stylatula, Acanthoptilum, Parazoanthus and Mnemiopsis. Depending on the organism these include one or more of the following: luciferyl sulfate, luciferase, and luciferin sulfokinase. These isolated components were found to be indistinguishable from those found in Renilla as evidenced by their reactivity in the Renilla bioluminescent system, by the spectral characteristics of the isolated luciferyl sulfates, by the molecular weights of the luciferases, and by the colors of the bioluminescence produced in vitro.
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  • 190
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    Journal of Cellular Physiology 81 (1973), S. 323-337 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In a microspectrophotometric study, photographic emulsions and a computer are used for measuring the hemoglobin content of a large number (about 50,000) of erythroid cells in fetal mice. Histograms of the hemoglobin content in erythroid cells illustrate the kinetics of erythropoiesis in yolk sac derived nucleated cells in the fetal peripheral blood, in fetal liver, and in fetal spleen. After the occasional extrusion of their nucleus, yolk sac derived erythrocytes remain as “macrocytes” in fetal circulation two or three days longer than the nucleated yolk sac derived erythrocytes do. Erythrocytes in fetal liver have a constant hemoglobin content of 28 pg 2 until day 17 of gestation. During further erythropoiesis in liver and then in the spleen, this amount is gradually adapted to the normal hemoglobin content in red blood cells of 16 pg.
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  • 191
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    Journal of Cellular Physiology 81 (1973), S. 365-369 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of lanthanum ions upon amoeboid movement and upon the redistribution of dense cytoplasmic inclusions following centrifugation were investigated in Amoeba discoides. Treatment with low lanthanum concentrations produced a characteristic inhibited condition in which locomotion ceased while saltatory movements continued. The resorting of stratified cytoplasm by local internal cytoplasmic flow was unaffected. This contrasted with p-hydroxymercuribenzoate treatment which inhibited both locomotion and saltatory movements. The inhibitory effects of lanthanum ions were antagonised by calcium ions but not by magnesium ions. It is reasoned that lanthanum competes for a peripheral binding site thereby disrupting a calcium-mediated regulatory mechanism of amoeboid movement.
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  • 192
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    Journal of Cellular Physiology 82 (1973) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 193
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    Journal of Cellular Physiology 82 (1973), S. 39-48 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: It is possible, by direct measurements, to demonstrate the presence of cyclic AMP in Tetrahymena. Its presence in a unicellular eukaryote suggests a fundamental role in cellular metabolism independent of its function as a “second messenger.” Caffeine, an inhibitor of phosphodiesterase, raises the intracellular concentration of cyclic AMP and causes an inhibition of cellular division. Dibutyryl cyclic AMP also causes a partial inhibition of proliferation and is potentiated by caffeine. Inhibited cells do not recover spontaneously during continued exposure to caffeine but the inhibition is reversible resulting in a synchronous wave of division peaking one generation time after return to fresh medium.Synchronous cells subjected to pulses of caffeine at various intervals of the cell cycle are set back in time as measured by the excess division delay. There is a linear relationship between the age of the cells at the pulse and the duration of the excess division delay such that the older the cells are the greater is the setback.Caffeine and dibutyryl cyclic AMP inhibit ciliary regeneration as does also deuterium oxide.
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  • 194
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    Journal of Cellular Physiology 82 (1973), S. 21-38 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Electrical properties of normal and dysgenic mouse skeletal muscle were studied by intracellular recording from embryonic cells developing in vitro. Passive membrane constants were determined from records of transmembrane potential responses to hyperpolarizing pulses of current using two types of analyses, assuming the tubes to be finite cylinders: the off transient and steady state analyses. The following properties of normal and dysgenic fibers were also studied. (a) membrane potentials (b) acetylcholine sensitivity (c) α-Bungarotoxin binding and (d) maximum rate of rise, overshoot and one-half fall time of the action potential. Rare electrotonic coupling between fibroblasts and myotubes was noted. An anomalous type of rectification Was observed in some fibers in which the transmembrane potential responses possessed under and overshoots. These responses may have affected the values of membrane constants as derived by the off transient analysis. In all parameters studied, including membrane constants derived by the steady state analysis, the cultured mouse cells resembled adult denervated mammalian muscle rather than innervated muscle. There were no differences between normal and dysgenic fibers with respect to any of the parameters studied. Dysgenic fibers did not contract although they displayed passive and active membrane properties like those in normal, non-dysgenic fibers.
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  • 195
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    Journal of Cellular Physiology 82 (1973), S. 133-150 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Metabolic inhibitors of RNA synthesis, such as actinomycin D and MPB (2-mercapto-1-(β-4-pyridethyl) benzimidazole) have often been used to test the possibility that transcription is required for some cellular process or to measure the „half-life“ of messenger RNA (mRNA). The basic assumption of this work has been that the primary effect of these inhibitors is on transcription alone, and that any effect on translation is secondary to the inhibition of mRNA synthesis. This assumption has been tested by examining the effects of these inhibitors at different doses on various parameters of the transcriptional and translation processes in mouse L-cells in culture, i.e., the rates of RNA, DNA, and protein synthesis; the level and size of cytoplasmic polysomes; the rates of polypeptide elongation and termination, and the role of peptide chain initiation in protein synthesis. The results indicate that the basic assumption is not correct since these inhibitors affect protein synthesis by inhibiting the rate of initiation and not the level of mRNA. Another implication of the experiments is that the average mRNA in L-cells has a half-life of at least one cell generation (16-18 hours), and that earlier values from experiments using actinomycin are underestimates. Cordycepin (3′-deoxyadenosine) which inhibits the production of ribosomal and messenger RNA also affects protein synthesis at the level of initiation. In contrast, two inhibitors of DNA synthesis (hydroxyurea and cytosine arabinoside) inhibit protein synthesis and the level of polysomes by some other mechanism.
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  • 196
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    Journal of Cellular Physiology 82 (1973), S. 189-198 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: SV101, the SV40-transformed subline of the mouse fibroblast line 3T3, is both serum- and density transformed, since it grows in both 1% and 10% calf serum, and grows beyond confluence in 10% calf serum. Negative selection at low cell density in 1% calf serum or in 10% agamma-depleted serum permits direct recovery of serum-revertant sublines of SV101. These sublines are unable to grow in 1% calf serum.Although negative selection at high cell density in 10% calf serum is known to permit recovery of density-revertant sublines of SV101, most density-revertants are not serum-revertant. However, all serum-revertants isolated so far are density-revertant as well.
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  • 197
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    Journal of Cellular Physiology 82 (1973), S. 231-238 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Variations in pH, serum concentration and the availability of Zn++ in the medium markedly influence the initiation of DNA synthesis in cultured chick embryo cells. This report considers the interplay of these factors with one another and with other factors such as type of medium, cell population density and the malignaut transformation in an attempt to better define the variables of the growth control system. Conditioned medium seems to protect the cells against the inhibitory effects of lowered pH. Increased serum concentration has a similar, but more striking effect. Increased serum concentration and pH, as well as decreased population density, which stimulate DNA synthesis, also lower the sensitivity of DNA synthesis to inhibition by Zn++ deprivation. Likewise, cell transformation by infection with Rous sarcoma virus lowers the sensitivity of DNA synthesis to inhibition by Zn++ deprivation and by pH reduction. The response of DNA synthesis to pH varies with the type and concentration of buffer used. It is concluded that there are a number of mutually interacting variables involved in the regulation of animal cell multiplication.
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  • 198
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    Journal of Cellular Physiology 82 (1973), S. 277-283 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effect of phytohemagglutinin (PHA) on the growth and number of granulocytic colonies (GC) developing on agar from bone marrow and spleen cells of normal and erythroleukemic mice inoculated with Rauscher leukemogenic virus was studied. Equal number of marrow cells from erythroleukemic mice produced twice as many colonies as those from normal mice. The number of GC developing from either normal and leukemic spleen cells was only 20% to 25% of that arising from marrow cells. The number of cells within each colony was significantly larger in GC formed by myelogenous leukemic cells than those arising from normal cells even though they had similar morphologic features. The addition of 100 μg of PHA per 105 cells reduced the number of GC arising from normal and leukemic cells by 35% and 50%, respectively. Treatment with periodate which mainly inhibits its mitogenic activity, abolished the inhibitory effects of PHA on proliferation of granulocytic cells.
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  • 199
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    Journal of Cellular Physiology 82 (1973), S. 339-347 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A temperature-sensitive (ts) mutant of Chinese hamster cells (K12) is blocked at a step late in G1, required for the initiation of DNA synthesis. The ts lesion is recessive in interspecific hybrids.
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